• Journal of Microbiology and Biotechnology
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• 제18권8호
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• pp.1470-1474
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• 2008

Nitrospira is a dominant member of nitrite-oxidizing bacteria (NOB) in nitrifying bioreactors as well as in natural habitats. In this study, Nitrospira NOB were investigated in the two nitrifying reactors operated with high and low dissolved oxygen (DO) concentrations for a period of 300 days. Phylogenetic and terminal restriction fragment length polymorphism analyses based on 168 rRNA gene sequences revealed that the Nitrospira community compositions of the two reactors during the early period related to group 1 and half of the Nitrospira community composition shifted to group 2 in the high-DO reactor after day 179, although there was no significant change in the low-DO reactor. These results suggested that DO was an important factor affecting Nitrospira community compositions in the nitrifying reactors.

• 약학회지
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• 제37권5호
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• pp.504-510
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• 1993

Human insulin gene is consisted of the polymorphic region with the repeating units, the regulatory sequence, the structural gene including the intervening sequence, and 3'-flanking region. The polymerase chain reaction, which amplifies the target DNA between two specific primers, has been performed for the amplification of human insulin gene and simple one-step cloning of it into Escherichia coli. Out of 1727 nuceotides compared, only 4 sites were variable: 5'-regulatory region(G2101$\rightarrow$AGG); IVS I(T2401$\rightarrow$A); Exon II(C2411 deletion); IVS II(A2740 dejection). The variations at the G2101 and T2401 were the same as those found in one American allele. The other two variations were observed only in the specific Korean allele. And, the enzyme digestion patterns among normal, insulin dependent diabetes mellitus, and non-insulin dependent diabetes mellitus were the same. On the other hand, PCR method showed the possibility of the quickaccess for the polymorphic region in terms of the restriction fragment length of polymorphism.

• Asian-Australasian Journal of Animal Sciences
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• 제27권10호
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• pp.1487-1492
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• 2014

This research applied and evaluated a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using cytochrome b gene to detect pork contamination in meatballs from local markets in Surabaya and Yogyakarta regions, Indonesia. To confirm the effectiveness and specificity of this fragment, thirty nine DNA samples from different meatball shops were isolated and amplified, and then the PCR amplicon was digested by BseDI restriction enzyme to detect the presence of pork in meatballs. BseDI restriction enzyme was able to cleave porcine cytochrome b gene into two fragments (131 bp and 228 bp). Testing the meatballs from the local market showed that nine of twenty meatball shops in Yogyakarta region were detected to have pork contamination, but there was no pork contamination in meatball shops in Surabaya region. In conclusion, specific PCR amplification of cytochrome b gen and cleaved by BseDI restriction enzymes seems to be a powerful technique for the identification of pork presence in meatball because of its simplicity, specificity and sensitivity. Furthermore, pork contamination intended for commercial products of sausage, nugget, steak and meat burger can be checked. The procedure is also much cheaper than other methods based on PCR, immunodiffusion and other techniques that need expensive equipment.

• 대한미생물학회지
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• 제34권6호
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• pp.561-571
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• 1999

Diagnosis of mycobacterial infection is dependent upon the isolation and identification of causative agents. The procedures involved are time consuming and technically demanding. To improve the laborious identification process mycobacterial systematics supported by gene analysis is feasible, being particularly useful for slowly growing or uncultivable mycobacteria. To complement genetic analysis for the differentiation and identification of mycobacterial species, an alternative marker gene, rpoB encoding the ${\beta}$ subunit of RNA polymerase, was investigated. rpoB DNAs (342 bp) were amplified from 52 reference strains of mycobacteria including Mycobacterium tuberculosis H37Rv (ATCC 27294) and clinical isolates by the PCR. The nucleotide sequences were directly determined (306 bp) and aligned using the multiple alignment algorithm in the MegAlign package (DNASTAR) and MEGA program. A phylogenetic tree was constructed with a neighborhood joining method. Comparative sequence analysis of rpoB DNA provided the basis for species differentiation. By being grouped into species-specific clusters with low sequence divergence among strains belonging to same species, all the clinical isolates could be easily identified. Furthermore RFLP analysis enabled rapid identification of clinical isolates.

• 대한두경부종양학회지
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• 제12권2호
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• pp.169-176
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• 1996

결핵의 진단은 특징적 병리조직양상, 항산균 염색에 의한 균 증명과 M. tuberculosis균 배양으로 이루어지나 형태적으로는 결핵이 의심되더라도 항산균이 조직표본이나 도말에서 검출되지 않거나 M. tuberculosis가 배양되지 않아 정확한 원인적 진단이 불가능할 경우가 많다. 이에 저자들은 경부 결핵성 임파선염으로 적출되어 보내어진 경부 임파선 조직의 신선한 조직이나 통상적으로 처리되어 제작된 파라핀 블록을 이용하여 M. tuberculosis에 특수한 반복성 DNA sequence인 IS986를 표적으로 한 primers을 사용하여 nested PCR방법을 이용하여 예민도가 높은 M. tuberculosis 검출로 빠른 시간 내에 결핵을 진단하고자 본 연구를 실시하였다. 최근 유전자 기술의 진보로 M. tuberculosis의 여러 항원들의 유전자가 클론화되고 그 염기 배열이 밝혀졌으며 이에 저자들은 결핵의 확진을 위하여 파라핀 포매조직을 대상으로 nested PCR에 의한 188bp의 DNA를 증폭한후 증폭한 DNA분절의 염기 배열을 결정한 후 Bst UI와 Hha I 효소를 이용한 소화과정을 거친 후 restriction fragment length polymorphism(RFLP)을 은염색에 의해 그 패턴에 의해 M. tuberculosis를 확인하고 또한 다른 종의 Mycobacteria를 배제시킬 수 있었다. 본 방법은 1$\sim$2일에 끝나며, 방사선물질을 사용하지 않으면서도 감도 및 특이성이 우수하여 일반 병리실힘실에서도 M. tuberculosis를 포함한 각종 항산균의 신속한 검출법으로 손쉽게 사용할 수 있다고 생각되어 이에 보고하였다.

• 과학수사학회지
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• 제11권4호
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• pp.276-282
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• 2017

사건현장에서 얻어질 수 있는 증거물로써 현대 분자수준의 수사기법에 힘입어 많은 미세증거물들의 가치가 확인되고 있다. 이러한 미세증거물에는 DNA처럼 개인 식별에 유용한 물질들도 포함되어 있지만, 실제 현장에서 개별적인 특성을 나타내는 증거물만을 수집하는 것은 쉽지 않기 때문에 아직 응용되지 못하거나 발굴되지 않은 증거물 후보군에 대해 연구가 지속되어야 할 필요가 있다. 본 연구에서는 16명의 사람으로부터 손에서 서식하는 세균 군집을 채취하였으며 미생물군집분석방법 중 하나인 제한효소 절편길이 다형성(T-RFLP) 기법을 개인 식별에 활용할 수 있는지의 여부를 조사하였다. 그 결과, 16개의 서로 다른 electropherogram을 얻을 수 있었고, Staphylococcus속과 Bacillus 속을 포함하여 개인마다 종류와 조성의 차이를 보이는 다양한 세균 분류군들이 손바닥에 서식하고 있음을 확인하였으며, 이를 개인을 둘러싼 환경조건과 성별 등의 요인들을 연관하여 해석하고자 하였다.

• 한국축산식품학회지
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• 제26권3호
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• pp.375-379
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• 2006

Food labeling regulations require that the meat species in various meat products are accurately declared to the consumer. Substitution or adulteration of costly meat with a cheaper one is one of the most common problems in the meat industry. In this study, PCR-restriction fragment length polymorphism(RFLP) method of the mitochondrial cytochrome b(mt cyt b) gene has been applied for identification of the origin of six mammalian meat species(beef, port horse, goat, mutton and deer) and three poultry meat species(chicken, turkey and duck) as raw materials for meat products. PCR was used to amplify a variable region of mt cyt b gene. Meat species differentiation was determined by digestion of the amplified products with a 359 bp fragment using HaeIII and HinfI restriction enzymes, which generated species-specific RFLP patterns. This PCR-RFLP DNA marker of mt cyt b gene could be very useful for the accurate and reliable identification and discrimination of animal meat species in routine analysis.

• 식물분류학회지
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• 제34권1호
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• pp.43-61
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• 2004

한국산 제비꽃속 32분류군과 일본산 2집단 등 총 34집단에 대한 유연관계를 알아보기 위하여 RAPD(randomly amplified polymorphic DNA), ISSR(inter simple sequence repeat) 및 PCR-RFLP(restriction fragment length polymorphism) 분석을 실시하였다. RAPD 분석에서는 40개의 primer 중 6개가 분류군 전체에서 반응을 보였고 이로부터 총 70개(98.6%)의 다형화 밴드를 얻었으며, ISSR 분석에서는 4개의 primer로 부터 28개(96.6%)의 다형화밴드를 얻었다. 엽록체 DNA의 non-coding부분을 이용한 PCR-RFLP 분석에서는 반응이 일어난 4지역에서 증폭된 약 6.78 kb의 DNA 각각에 대하여 15가지 제한효소를 처리한 결과 총 80개의 restriction site를 얻었으며 그중 16 site는 polymorphic하게 나타나 20%의 다형화를 보였다. 본 연구에서 다룬 3가지 형질에 의한 유집분석 결과 각각의 형질에 의해서는 서로 일치하지 않는 유집형태를 보였지만 3가지 형질을 통합한 결과는 진정제비꽃절(sect. Nomimium)내 아절과 계열간 구분이 명확하게 나타났으며 무경종과 유경종도 구별이 가능하여 외부형태형질에 의한 기존의 분류체계와 일치하였다. 그러나 노랑제비꽃절(sect. Chamaemelanium)은 진정제비꽃절과 독립적인 군을 형성하지 않고 진정제비꽃절 내 무경종그룹인 Patellares아절과 Vaginatae아절 사이에 위치하여 나타났다. 형태적인 변이가 매우 심한 분류군으로 알려진 태백제비꽃군(V. albida complex)은 Patellares아절 내에서 하나의 군으로 유집되어 Pinnatae계열로 처리하는 것이 타당할 것으로 생각된다. 본 연구에서 사용한 3가지 형질 중 RAPD 분석방법은 ISSR과 PCR-RFLP 분석보다 제비꽃속의 종간 유연관계를 밝히는데 더 유용한 것으로 판단된다.

• International Journal of Oral Biology
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• 제31권3호
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• pp.107-112
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• 2006

Aldehyde dehydrogenase (ALDH) plays an important role in alcohol metabolism; ALDH is responsible for the oxidation of acetaldehyde generated during alcohol oxidation. ALDH is also known to oxidize various other endogenous and exogenous aldehydes. Cytochrome P-450 2E1 (CYP2E1), a liver microsomal enzyme, also metabolizes acetaldehyde and ethanol and can be induced by other inducers including acetone and ethanol. We examined single nucleotide polymorphisms (SNP) of ALDH and CYP2E1 genotypes in Korean. Restriction fragment length polymorphism (RFLP) method was used to determine ALDH and CYP2E1 SNP. Mutation in ALDH was 60% (heterozygote 46.7% and homozygote 13.3%) among 15 cases. CYP2E1 mutation was 52.7% (heterozygote 47.4% and homozygote 5.3%) among 19 cases.

• 한국동물위생학회지
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• 제31권3호
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• pp.369-374
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• 2008

The objective of this study was to differentiate the beef species between Hanwoo and Holstein from a total of 1,081 beef samples using PCR-RFLP of MC1R gene. When a PCR product of 403 bp specific band amplified from bovine MC1R gene sequence was digested with restriction enzyme MspA1I, Hanwoo type showed 2 bands, 220 bp and 183 bp size bands. Holstein type, however, showed three bands, 220 bp, 138 bp and 45 bp size band, respectively. The results of the differential test for beef species were as following; 7 samples (0.64%) were determined to Holstein type, of which 4 were submitted from administrative authorities, other 3 from self-collection planing, and none from civilian clients including school.